Journal
FEBS LETTERS
Volume 588, Issue 17, Pages 2964-2970Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.febslet.2014.05.050
Keywords
Holophytochrome; Chromophore; Phytochromobilin; Heme oxygenase; Bilin reductase
Funding
- Next-Generation BioGreen 21 Program
- Rural Development Administration, Republic of Korea [PJ00949101]
- Basic Science Research Program through the National Research Foundation of Korea (NRF) - Ministry of Education, Science and Technology [2011-0012362, 2012R1A1A2000706]
- National Research Foundation of Korea [2012R1A1A2000706, 2011-0012362] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
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We have successfully developed a system to produce full-length plant phytochrome assembled with phytochromobilin in Pichia pastoris by co-expressing apophytochromes and chromophore biosynthetic genes, heme oxygenase (HY1) and phytochromobilin synthase (HY2) from Arabidopsis. Affinity-purified phytochrome proteins from Pichia cells displayed zinc fluorescence indicating chromophore attachment. Spectroscopic analyses showed absorbance maximum peaks identical to in vitro reconstituted phytochromobilin-assembled phytochromes, suggesting that the co-expression system is effective to generate holo-phytochromes. Moreover, mitochondria localization of the phytochromobilin biosynthetic genes increased the efficiency of holophytochrome biosynthesis. Therefore, this system provides an excellent source of holophytochromes, including oat phytochrome A and Arabidopsis phytochrome B. (C) 2014 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.
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